Can further enzymatic reactions be performed in the µ Column, such as cDNA labeling?
With MACS® Technology, many kinds of enzymatic reactions can be performed directly in the column, allowing a fast workflow and minimized loss of sample, as no tube-to-tube transfer of the sample is necessary. Furthermore, the sample purification after the reaction is easily performed without additional phenol extraction or precipitation. DNase I treatment When processing cell lysate to mRNA and first-strand cDNA, an in-column genomic DNA removal with DNAse I can optionally be performed. As the µMACS™ mRNA isolation yields mRNA virtually free of genomic DNA contamination, this step is only recommended for very limited samples in combination with highly sensitive downstream applications such as real-time PCR from mRNA isolated from a few cells only, when intron-spanning primers are not available. RNase H treatment Although residual mRNA does generally not interfere with downstream PCR or real-time PCR, RNAse H removal of mRNA in mRNA-cDNA hybrids can be performed directly in the column.
